Larrea F, Méndez I, Parra A, Espinosa de los Monteros A
Department of Reproductive Biology, Instituto Nacional de la Nutrición Salvador Zubirán, México D.F.
Hum Reprod. 1993 Oct;8(10):1617-22. doi: 10.1093/oxfordjournals.humrep.a137901.
In this study, the molecular heterogeneity of prolactin was analysed in serum from normal women throughout pregnancy. Lectin affinity chromatography and denaturing polyacrylamide gel electrophoresis under reducing and non-reducing conditions, followed by Western blotting and immunostaining were used to resolve and identify the molecular variants of prolactin. During the first trimester, large molecular forms (64 and 53 kDa) and those corresponding to glycosylated and non-glycosylated prolactin (25 and 23 kDa, respectively) were present either under reducing or non-reducing conditions. The 64 and 23 kDa were the predominant species at this stage of gestation. As pregnancy progressed, the 64 kDa variant, which did not bind to concanavalin A, decreased until disappeared at the third trimester of gestation. The unbound/bound ratio of serum prolactin to concanavalin A increased only at the third trimester; however, the relative proportions of concanavalin A-bound prolactin did not show statistically significant changes along the gestational period. The results demonstrated the occurrence of changes in the heterogeneity of prolactin during gestation and further confirmed previous observations that various forms of non-glycosylated prolactin are indeed the predominant species in serum from normal women throughout pregnancy.
在本研究中,对正常女性孕期血清中催乳素的分子异质性进行了分析。采用凝集素亲和层析以及在还原和非还原条件下的变性聚丙烯酰胺凝胶电泳,随后进行蛋白质印迹和免疫染色,以解析和鉴定催乳素的分子变体。在孕早期,无论是在还原还是非还原条件下,均存在大分子形式(64 kDa和53 kDa)以及对应糖基化和非糖基化催乳素的形式(分别为25 kDa和23 kDa)。64 kDa和23 kDa的变体在妊娠此阶段占主导。随着妊娠进展,不与伴刀豆球蛋白A结合的64 kDa变体减少,直至在妊娠晚期消失。血清催乳素与伴刀豆球蛋白A的未结合/结合比率仅在妊娠晚期增加;然而,伴刀豆球蛋白A结合的催乳素的相对比例在整个妊娠期未显示出统计学上的显著变化。结果表明妊娠期催乳素的异质性发生了变化,并进一步证实了先前的观察结果,即各种形式的非糖基化催乳素确实是正常女性孕期血清中的主要形式。